Development of a novel alternative to animal testing for the evaluation of respiratory allergic sensitizing potential of chemicals

• Project/Area Number: 18K11658
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 63030:Chemical substance influence on environment-related
• Research Institution: Tokyo Medical University
• Principal Investigator: Mizoguchi Izuru 東京医科大学, 医学部, 講師 (00569527)
• Co-Investigator(Kenkyū-buntansha): 善本 隆之 東京医科大学, 医学部, 教授 (80202406)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 感作性評価法 / 動物実験代替法 / 呼吸器感作性 / 皮膚感作性 / Th2反応 / 感作性 / アレルギー / 化学物質 / 3次元共培養系 / 樹状細胞

Outline of Final Research Achievements

Recently, we established a new 3-dimentional (3D) dendritic cell (DC) coculture system consisting of human airway epithelial cell line, immature DCs derived from human peripheral monocytes, and lung fibroblast cell line. This coculture system was shown to successfully discriminate respiratory sensitizers from skin sensitizers using 6 representative chemical sensitizers by more enhanced mRNA expression of key costimulatory molecule OX40 ligand, which is important for Th2 differentiation, in DCs. Herein, we have further established a new 2-step 3D DC/T coculture system by introducing T cells in the DC coculture system, in which IL-4 expression in T cells, that are the Key event 4 in the adverse outcome pathway of sensitization, can be used as a marker to discriminate these sensitizers. To increase the versatility, we are also trying to generate DC progenitor cell lines and T cell lines to apply for it.

Academic Significance and Societal Importance of the Research Achievements

化学物質の感作性試験法には、近年、動物を使わない代替法の開発が急務となっている。感作性化学物質には、呼吸器と皮膚感作性の2種類あり、両者に対して講ずるべき危機管理対策のレベルが全く異なるにも関わらず、既存のin vitro感作性評価法では、両者を見分けることができない。また、これまでに、感作性の有害性発現経路のKey event 4のT細胞を指標にした代替試験法は開発されていない。本研究により、T細胞でのIL-4発現を指標に呼吸器と皮膚感作性を識別する樹状細胞とT細胞の3次元共培養系の開発の可能性が示唆され、さらに汎用性の高い系ができれば、産業界への影響は多大でその社会的意義も大きい。

Research Products

• [Journal Article] IL-23p19 and CD5 antigen-like form a possible heterodimeric cytokine and contribute to experimental autoimmune encephalomyelitis development. (2021)
  • Author(s): Hasegawa H, Mizoguchi I, Orii N, Inoue S, Katahira Y, Yonteo T, Mingli X, Miyazaki T, Yoshimoto T
  • Journal Title: Sci Rep. Volume: 11(1) Issue: 1 Pages: 5266-5266
  • DOI: 10.1038/s41598-021-84624-9
  • Peer Reviewed / Open Access
• [Journal Article] Interleukin-1β in peripheral monocytes is associated with seizure frequency in pediatric drug-resistant epilepsy. (2021)
  • Author(s): Yamanaka G, Takamatsu T, Morichi S, Yamazaki T, Mizoguchi I, Ohno K, Watanabe Y, Ishida Y, Oana S, Suzuki S, Kashiwagi Y, Takata F, Sakuma H, Yoshimoto T, Kato M, Kawashima H
  • Journal Title: J Neuroimmunol. Volume: 352 Pages: 577475-577475
  • DOI: 10.1016/j.jneuroim.2021.577475
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Microbial antigen-presenting extracellular vesicles derived from genetically modified tumor cells promote antitumor activity of dendritic cells (2021)
  • Author(s): Tomoko Ito, Kikuya Sugiura, Aya Hasegawa, Wakana Ouchi, Takayuki Yoshimoto, Izuru Mizoguchi, Toshio Inaba, Katsuyuki Hamada, Masazumi Eriguchi, Yoshiyuki Koyama
  • Journal Title: Pharmaceutics Volume: 13 Issue: 1 Pages: 57-69
  • DOI: 10.3390/pharmaceutics13010057
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] EBV-induced gene 3 augments IL-23Rα protein expression through a chaperone calnexin. (2020)
  • Author(s): Mizoguchi I, Ohashi M, Hasegawa H, Chiba Y, Orii N, Inoue S, Kawana C, Xu M, Sudo K, Fujita K, Kuroda M, Hashimoto S, Matsushima K, and Yoshimoto T.
  • Journal Title: J Clin Invest. Volume: 130(11) Issue: 11 Pages: 6124-6140
  • DOI: 10.1172/jci122732
  • NAID: 120007052824
  • Peer Reviewed / Open Access
• [Journal Article] IL-12ヘテロダイマーサイトカインファミリーIL-12、IL-23、IL-35 (2020)
  • Author(s): 片平泰弘、井上槙也、溝口 出、善本隆之
  • Journal Title: 医学のあゆみ「サイトカインのすべて」 Volume: 別冊 Pages: 53-60
• [Journal Article] Necroptosis of intestinal epithelial cells induces type 3 innate lymphoid cell-dependent lethal ileitis. (2019)
  • Author(s): Shindo R, Ohmuraya M, Komazawa-Sakon S, Miyake S, Deguchi Y, Yamazaki S, Nishina T, Yoshimoto T, Kakuta S, Koike M, Uchiyama Y, Konishi H, Kiyama H, Mikami T, Moriwaki K, Araki K, Nakano H.
  • Journal Title: iScience Volume: 15 Pages: 536-551
  • DOI: 10.1016/j.isci.2019.05.011
  • Peer Reviewed / Open Access
• [Journal Article] Plasmacytoid dendritic cells protect against immune-mediated acute liver injury via IL-35. (2019)
  • Author(s): Koda Y, Nakamoto N, Chu P, Ugamura A, Teratani T, Shiba S, Taniki N, Sujino T, Miyamoto K, Mikami Y, Suzuki T, Yamaguchi A, Morikawa R, Sato K, Yoshimoto T, Kanai K.
  • Journal Title: J Clin Invest Volume: 129(8) Issue: 8 Pages: 3201-3213
  • DOI: 10.1172/jci125863
  • Peer Reviewed / Open Access
• [Journal Article] Two cases of extremely rare primary pure squamous cell carcinoma of the breast. (2018)
  • Author(s): Yoneto T, Hasumi K, Yoshimoto T, Takahashi N, Takeda Y.
  • Journal Title: Medicine (Baltimore) Volume: 97(37) Issue: 37 Pages: e12340-e12340
  • DOI: 10.1097/md.0000000000012340
  • Peer Reviewed / Open Access
• [Journal Article] Protective effects against tumors and infection by IL-27 through promotion of expansion and differentiation of hematopoietic stem cells into myeloid progenitors. (2018)
  • Author(s): Orii N, Mizoguchi I, Chiba Y, Hasegawa H, Ohashi M, Xu M, Nagai T, Ochiai M, Mochizuki Y, Owaki T, Yoshimoto T.
  • Journal Title: Oncoimmunology Volume: - Issue: 5 Pages: e1421892-e1421892
  • DOI: 10.1080/2162402x.2017.1421892
  • Peer Reviewed
• [Journal Article] Regulation of myelopoiesis by proinflammatory cytokines in infectious diseases. (2018)
  • Author(s): Chiba Y, Mizoguchi I, Hasegawa H, Ohashi M, Orii N, Nagai T, Sugahara M, Miyamoto Y, Xu M, Owaki T, Yoshimoto T.
  • Journal Title: Cell. Mol. Life Sci. Volume: 75 Issue: 8 Pages: 1363-1376
  • DOI: 10.1007/s00018-017-2724-5
  • Peer Reviewed / Open Access
• [Journal Article] Interleukin-27 exerts its antitumor effects by promoting differentiation of hematopoietic stem cells to M1 macrophages. (2018)
  • Author(s): Chiba Y, Mizoguchi I, Furusawa J, Hasegawa H, Ohashi M, Xu M, Owaki T, Yoshimoto T.
  • Journal Title: Cancer Res. Volume: 78 Issue: 1 Pages: 182-194
  • DOI: 10.1158/0008-5472.can-17-0960
  • Peer Reviewed / Open Access
• [Presentation] アレルギー感作性を評価する新規動物実験代替法の開発 (2020)
  • Author(s): 溝口 出、川名千晶、長谷川英哲、折井直子、井上槙也、村上史浩、米戸敏彦、徐 明利、善本隆之
  • Organizer: 東京医科大学記念館ポスター発表懇談会
• [Presentation] 活性化T細胞から産生されるIL-23p19サブユニットの役割 (2020)
  • Author(s): 長谷川英哲、溝口 出、折井直子、川名千晶、井上槙也、村上史浩、米戸敏彦、徐 明利、善本隆之
  • Organizer: 東京医科大学記念館ポスター発表懇談会
• [Presentation] ESAT-6抗原提示エクソソームによる免疫活性化 (2020)
  • Author(s): 小山義之、伊藤智子、江里口正純、善本隆之、溝口 出、杉浦喜久弥、長谷川 綾、大内若菜、稲葉俊夫
  • Organizer: 第7回日本細胞外小胞学会学術集会
• [Presentation] 美白成分による自己免疫性白斑症誘発の作用機序の解明 (2020)
  • Author(s): 坂本恵梨、片平泰弘、溝口 出、井上槙也、長谷川英哲、善本隆之
  • Organizer: 第186回東京医科大学医学会総会
• [Presentation] 化粧品美白成分による白斑症誘発の作用機序の解明とそのin vitro評価法の開発 (2020)
  • Author(s): 坂本恵梨、片平泰弘、井上槙也、古阪悠馬、渡邉有麻、長谷川英哲、米戸敏彦、徐 明利、溝口 出、善本隆之
  • Organizer: 第33回日本動物実験代替法学会
• [Presentation] アレルギー感作性・誘発性を評価する新規動物実験代替法の開発 (2019)
  • Author(s): 溝口 出、川名千晶、井上槙也、折井直子、長谷川英哲、米戸敏彦、徐 明利、善本隆之
  • Organizer: 第8回医薬工3大学包括連携推進シンポジウム
• [Presentation] 化学物質のアレルギー感作性・誘発性を評価する新規動物実験代替法の開発 (2019)
  • Author(s): 溝口 出、川名千晶、井上槙也、折井直子、長谷川英哲、米戸敏彦、徐 明利、善本隆之
  • Organizer: 第32回日本動物実験代替法学会
• [Presentation] Development of a novel alternative method for evaluation of sensitizing potential and allergenicity by measuring human T cell activation and differentiation in vitro. (2019)
  • Author(s): Mizoguchi I, Inoue S, Orii N, Hasegawa H, Xu X, Yoshimoto T.
  • Organizer: 第48回日本免疫学会総会・学術集会
• [Presentation] Prediction of chemical respiratory and contact sensitizers by OX40L expression in dendritic cells using a novel 3D co-culture system. (2018)
  • Author(s): Ohashi, M., Mizoguchi, I., Chiba, Y., Hasegawa, H., Orii, N., Kan, S., Xu, M., Ochiai, N., Owaki, T., and Yoshimoto, T.
  • Organizer: 東京医科大学記念館ポスター発表懇談会
• [Presentation] 新しい3次元共培養系を用いた呼吸器感作性代替法の開発 (2018)
  • Author(s): 善本隆之、大橋美緒、長谷川英哲、折井直子、徐明利、大脇敏之、溝口 出
  • Organizer: 第45回日本毒性学会学術年会
  • Invited
• [Presentation] 化学物質の呼吸器感作性in vitro評価法の開発 (2018)
  • Author(s): 溝口 出、折井直子、長谷川英哲、善本隆之
  • Organizer: 第6期LRI研究報告会
• [Presentation] Prediction of chemical respiratory sensitizers by OX40L expression in dendritic cells using a new 3D co-culture system. (2018)
  • Author(s): Yoshimoto, T., Hasegawa, H., Naoko, O., Xu, M., and Mizoguchi, I.
  • Organizer: The 4th International Conference on Toxicity Testing Alternatives & Translational Toxicology and the 2nd Asian Congress on Alternatives.